Intra-patient genomic heterogeneity of single circulating tumor cells (ctcs) associated to phenotypic ctc heterogeneity in metastatic castrate resistant prostate cancer (mcrpc)

Abstract

The disclosure provides methods correlating intra-patient genomic heterogeneity of single CTCs with phenotypic heterogeneity in each of a population of prostate cancer (PCa) patients.

What is claimed is: 
     
         1 . A method for correlating intra-patient genomic heterogeneity of single CTCs with phenotypic heterogeneity in a prostate cancer (PCa) patient comprising:
 (a) performing a direct analysis comprising immunofluorescent staining and morphological characterization of nucleated cells in a blood sample obtained from the patient to identify and enumerate circulating tumor cells (CTC);   (b) isolating the CTCs from said sample;   (c) individually characterizing genomic alterations and phenotypic features to generate a profile for each of the CTCs, and   (d) correlating genomic heterogeneity of single CTCs with phenotypic heterogeneity in the PCa patient.   
     
     
         2 . A method for correlating intra-patient genomic heterogeneity of single CTCs with phenotypic heterogeneity in each of a population of prostate cancer (PCa) patients comprising:
 (a) performing a direct analysis comprising immunofluorescent staining and morphological characterization of nucleated cells in a blood sample obtained from the patient to identify and enumerate circulating tumor cells (CTC);   (b) isolating the CTCs from said sample;   (c) individually characterizing genomic alterations and phenotypic features to generate a profile for each of the CTCs;   (d) correlating individual genomic heterogeneity of single CTCs with phenotypic heterogeneity in each of the population of PCa patients, and   (e) analyzing said correlations of individual genomic heterogeneity of single CTCs with phenotypic heterogeneity across the population of PCa patients to identify a universal correlation of individual genomic heterogeneity of single CTCs with phenotypic heterogeneity.   
     
     
         3 . The method of  claim 1 , wherein said population of prostate cancer are similarly situated with regard to one or more patient demographics. 
     
     
         4 . The method of  claim 1 , wherein said demographics comprise therapy or line of therapy. 
     
     
         5 . The method of  claim 4 , wherein therapy is hormone directed therapy or chemotherapy. 
     
     
         6 . The method of  claim 5 , wherein said hormone directed therapy comprises Androgen Deprivation Therapy (ADT). 
     
     
         7 . The method of  claim 6 , wherein said ADT is a second line hormonal therapy. 
     
     
         8 . The method of  claim 7 , wherein said second line hormonal therapy blocks synthesis of androgen or inhibits Androgen Receptor (AR). 
     
     
         9 . The method of  claim 8 , wherein said second line hormonal therapy is selected from the group consisting of abiraterone acetate, ketoconazole and aminoglutethimide. 
     
     
         10 . The method of  claim 5 , wherein said chemotherapy is taxane therapy. 
     
     
         11 . The method of  claim 1  or  2 , wherein the immunofluorescent staining of nucleated cells comprises pan cytokeratin, cluster of differentiation (CD) 45, diamidino-2-phenylindole (DAPI) and androgen receptor (AR). 
     
     
         12 . The method of  claim 1  or  2 , wherein said morphological characterization comprises determination of one or more of the group consisting of nucleus size, nucleus shape, presence of holes in nucleus, cell size, cell shape and nuclear to cytoplasmic ratio, nuclear detail, nuclear contour, prevalence of nucleoli, quality of cytoplasm and quantity of cytoplasm. 
     
     
         13 . The method of  claim 1  or  2 , wherein said phenotypic features are selected from the group listed in  FIG. 3  D. 
     
     
         14 . The method of  claim 1  or  2 , wherein said genomic alterations are copy number variation (CNV) alterations. 
     
     
         15 . The method of  claim 14 , wherein said CNV alterations are selected from the group listed in  FIG. 3  D. 
     
     
         16 . The method of  claim 2 , wherein said universal correlation is used to identify a phenotypic profile that corresponds to a genotypic profile. 
     
     
         17 . The method of  claim 16 , wherein identification of said phenotypic profile obviates the need for characterizing said genomic alterations. 
     
     
         18 . The method of  claim 17 , wherein said phenotypic profile is capable of predicting emergence of resistant disease. 
     
     
         19 . The method of  claim 18 , comprising resistance to hormone directed therapy or chemotherapy. 
     
     
         20 . The method of  claim 19 , wherein said hormone directed therapy comprises Androgen Deprivation Therapy (ADT). 
     
     
         21 . The method of  claim 20 , wherein said ADT is a second line hormonal therapy. 
     
     
         22 . The method of  claim 21 , wherein said second line hormonal therapy blocks synthesis of androgen or inhibits Androgen Receptor (AR).